17 Questions
What is the purpose of adding proteinase K in DNA extraction?
To break down peptide bonds and digest contaminating proteins
Why is salt added during DNA extraction?
To stabilize the negative charged phosphate groups in DNA
How is DNA separated from other molecules in the extraction process?
By precipitating DNA out of the solution with ice-cold alcohol
What is the function of sodium dodecyl sulfate (SDS) in DNA isolation?
To solubilize proteins and lipids in cell membranes
What happens to the DNA after white precipitate forms in the extraction process?
It is washed and resuspended
What is the purpose of agarose gel electrophoresis?
Separate DNA molecules by size
Why is ethidium bromide added to the agarose solution in gel electrophoresis?
To stain the gel and visualize DNA bands
Which component of the loading buffer helps in monitoring migrating DNA bands?
Dyes
How does the size of DNA molecules affect their migration in agarose gel electrophoresis?
Smaller DNA molecules migrate more slowly
Why is a comb inserted into the gel cast during agarose gel preparation?
To create wells for loading DNA samples
What role does agarose concentration play in agarose gel electrophoresis?
Affects the speed of DNA migration
What is the purpose of keeping the restriction enzyme on ice or in a thermoresistant container?
To prevent the enzyme from being denatured
How many microfuge tubes are typically used in setting up a restriction enzyme digest?
1
What is the purpose of incubating the reaction mixture at 65°C?
To heat and activate the restriction enzyme
What is 'star activity' in the context of restriction enzyme digestion?
Cutting at sites similar but distinct from typical digestion sites
Why is it important to run DNA in an agarose gel after a restriction digest?
To ensure that the digest was successful
In what scenario would multiple enzymes need to be used for a restriction digest?
When encountering incompatible buffer conditions or temperatures
Test your knowledge on DNA extraction, purification, and analysis techniques. Learn about the process of lysing cells, using detergents for membrane dissolution, and the role of Proteinase K in isolating DNA. Prepare for understanding restriction enzyme analysis and other related topics.
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